Interaction Details


Details of experiments between Q92934 and P62258
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: BAD
Fragment: 113-122
Site: 14-3-3_pSer118
Modification: (SEP)118
Sequence: ELRRMSDEFV
Construct: ELRRM(pSer)DEFV (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.22
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 116
P value: 1.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 2

Protein A

Protein: BAD
Fragment: 113-122
Site: 14-3-3_pSer118
Modification: (SEP)118
Sequence: ELRRMSDEFV
Construct: ELRRM(pSer)DEFV (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.05
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 0.59
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 3

Protein A

Protein: BAD
Fragment: 119-128
Site: 14-3-3_pSer124
Modification: (SEP)124
Sequence: DEFVDSFKKG
Construct: DEFVD(pSer)FKKG (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.05
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 2.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 4

Protein A

Protein: BAD
Fragment: 119-128
Site: 14-3-3_pSer124
Modification: (SEP)124
Sequence: DEFVDSFKKG
Construct: DEFVD(pSer)FKKG (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.01
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 0.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 5

Protein A

Protein: BAD
Fragment: 129-138
Site: 14-3-3_pSer134
Modification: (SEP)134
Sequence: LPRPKSAGTA
Construct: LPRPK(pSer)AGTA (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.41
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 116
P value: 3.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.77
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 6

Protein A

Protein: BAD
Fragment: 129-138
Site: 14-3-3_pSer134
Modification: (SEP)134
Sequence: LPRPKSAGTA
Construct: LPRPK(pSer)AGTA (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.27
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 143
P value: 2.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.42
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 7

Protein A

Protein: BAD
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: AVEIRSRHSS
Construct: AVEIR(pSer)RHSS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.11
Holdup BI standard deviation: 0.42
Immobilized partner concentration (10-6M): 116
P value: 0.2
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 8

Protein A

Protein: BAD
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: AVEIRSRHSS
Construct: AVEIR(pSer)RHSS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.07
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 143
P value: 0.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 9

Protein A

Protein: BAD
Fragment: 70-79
Site: 14-3-3_pSer75
Modification: (SEP)75
Sequence: RSRHSSYPAG
Construct: RSRHS(pSer)YPAG (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.07
Holdup BI standard deviation: 0.51
Immobilized partner concentration (10-6M): 143
P value: 0.08
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 10

Protein A

Protein: BAD
Fragment: 94-103
Site: 14-3-3_pSer99
Modification: (SEP)99
Sequence: RGRSRSAPPN
Construct: RGRSR(pSer)APPN (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.44
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 5.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.83
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))